Cellular assay to study β-arrestin recruitment by the cannabinoid receptors 1 and 2

Bouma, J.; Soethoudt, M.; Gils, N. van; Xia, L.; Stelt, M, van der; Heitman, L.H. (2022)

Part of book or chapter of book

Cannabinoid receptor 1 (CB₁R) and cannabinoid receptor 2 (CB₂R) are G protein-coupled receptors (GPCRs) that activate a variety of pathways upon activation by (partial) agonists including the G protein pathway and the recruitment of β-arrestins. Differences in the activation level of these pathways lead to biased signaling. Here, we describe a detailed protocol to characterize the potency and efficacy of ligands to induce or inhibit β-arrestin recruitment to the human CB₁R and CB₂R using the PathHunter^® assay. This is a cellular assay that uses a β-galactosidase complementation system which has a chemiluminescent read-out and can be performed in 384-well plates. We have successfully used this assay to characterize a set of reference ligands (both agonists, antagonists, and an inverse agonist) on human CB₁R and CB₂R, of which some examples will be presented here.

• All authors: Bouma, J.; Soethoudt, M.; Gils, N. van; Xia, L.; Stelt, M, van der; Heitman, L.H.
• Editor(s): Maccarrone, M.
• Date: 2022-09-25
• Title of host publication: Methods in Molecular Biology; Endocannabinoid Signaling
• Volume: 2576
• Pages: 189 - 199
• ISBN (print): 9781071627273
• ISBN (electronic): 9781071627280
• DOI: doi:10.1007/978-1-0716-2728-0_15
• Name: 2576